黄芪甲苷纳米结构脂质载体的制备及其对肺癌作用的体内外研究

Preparation of astragaloside-loaded nanostructured lipid carriers and their effects on lung cancer in vitro and in vivo

目的制备黄芪甲苷(AST)纳米结构脂质载体(AST-NLCs),探讨AST对髓源抑制性细胞(MDSCs)和A549肺癌细胞的生长抑制作用及对荷瘤鼠的抗肿瘤药效。方法采用高压均质法,以粒径和多分散指数(PDI)为指标进行单因素实验,考察分散功率、分散时间、均质压力以及均质次数对AST-NLCs粒径的影响从而筛选得到最佳工艺条件,同时评价ASTNLCs的体外溶出行为。采用CCK-8法比较AST和AST-NLCs对MDSCs和A549细胞的抑制作用。建立A549 BALB/c荷瘤小鼠模型,研究AST-NLCs对A549荷瘤小鼠的体内抗肿瘤作用。最后通过病理切片检查,考察制剂的静脉刺激性。结果优化所得AST-NLCs的粒径为(136.7±5.2)nm,PDI为(0.162±0.009),Zeta电位为(-21.2±0.7)mV,包封率和载药量分别达到(83.5±1.57)%和(12.8±0.89)%。同AST原药粉比较,AST-NLCs的体外溶出明显提高。CCK-8测定结果显示AST-NLCs对MDSCs和A549细胞生长具有显著地抑制作用,在A549荷瘤鼠的体内实验中,AST-NLCs的抑瘤效果也显著高于AST组。组织病理切片的观察结果显示,AST组存在明显的静脉刺激性,NLCs能够降低AST静脉刺激性。结论高压均质法制备的AST-NLCs粒径大小分布均匀,药物包封率高,能显著提高药物的体外溶出度且具有明显的缓释效果,在血浆中能稳定存在。AST-NLCs能够通过抑制MDSCs和A549细胞达到抗肿瘤作用,抑瘤效果优于AST,且降低了AST的静脉刺激性。

Objective To prepare astragaloside-loaded nanostructured lipid carriers(AST-NLCs)and investigate their inhibitory effect on the growth of myeloid derived suppressor cells(MDSCs)and lung cancer A549 cells and the anti-tumor effect on tumor-bearing mice.Methods The hot melting high-pressure homogenization method was used to conduct single-factor experiments with particle size and polydispersity index(PDI)as indicators to determine the effect of stirring power,stirring time,homogeneous pressure and cycle times on the particle size to screen the best process conditions.The dissolution behavior of AST-NLCs was evaluated in vitro.The effects of AST and AST-NLCs on MDSCs and A549 cells were compared by CCK-8 assay.A549 BALB/c tumor-bearing mice model was established and the anti-tumor effect of AST-NLCs on A549 tumor-bearing mice in vivo was determine.Finally,the venous irritation of the preparation was measured by pathological section examination.Results The optimized particle size of AST-NLCs was(136.7±5.2)nm,the PDI was(0.162±0.009),the Zeta potential was(-21.2±0.7)mV,and the entrapment efficiency and drug loading were(83.5±1.57)%and(12.8±0.89)%,respectively.As compared with AST powder,AST-NLCs improved drug dissolution in vitro significantly.CCK-8 data showed that AST-NLCs significantly inhibited the growth of MDSCs and A549 cells.In the in vivo experiments on A549 tumor-bearing mice,the antitumor effect of AST-NLCs was also significantly higher than that of AST.Histopathological section showed that the AST group had obvious venous irritation,but NLCs reduced the AST venous irritation.Conclusion The particle size of GPS-NLCs prepared by high pressure homogenization is uniform,with high drug encapsulation efficiency.The in vitro dissolution rate of the drug is significantly improved with obvious sustained release effect.It is stable in the plasma.AST-NLCs achieve the anti-tumor effect via inhibiting MDSCs and A549 cells and the effect is better than that of AST.It also reduces the venous irritation of AST.